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Numerous cancers, including prostate cancer (PCa), are addicted to transcription programs driven by specific genomic regions known as super-enhancers (SEs). The robust transcription of genes at such SEs is enabled by the formation of phase-separated condensates by transcription factors and coactivators with intrinsically disordered regions. The androgen receptor (AR), the main oncogenic driver in PCa, contains large disordered regions and is co-recruited with the transcriptional coactivator mediator complex subunit 1 (MED1) to SEs in androgen-dependent PCa cells, thereby promoting oncogenic transcriptional programs. In this work, we reveal that full-length AR forms foci with liquid-like properties in different PCa models. We demonstrate that foci formation correlates with AR transcriptional activity, as this activity can be modulated by changing cellular foci content chemically or by silencing MED1. AR ability to phase separate was also validated in vitro by using recombinant full-length AR protein. We also demonstrate that AR antagonists, which suppress transcriptional activity by targeting key regions for homotypic or heterotypic interactions of this receptor, hinder foci formation in PCa cells and phase separation in vitro. Our results suggest that enhanced compartmentalization of AR and coactivators may play an important role in the activation of oncogenic transcription programs in androgen-dependent PCa.

Cognitive-behavioural models of auditory verbal hallucinations (voices) predict that the interpretation of voices determines the levels of distress experienced by voice-hearers. Examining the contribution of these voice appraisals is central to the delivery of effective psychological interventions for the management of distressing psychotic symptoms. This meta-analysis synthesised evidence from studies that tested the relationship between a range of appraisals and several distress measures (voice-related and emotional distress) in individuals with schizophrenia-spectrum diagnoses. A database search (PsycINFO, PubMed and Web of Science) was conducted for articles published up to August 2020. Twenty-eight eligible studies, comprising of 1497 clinical participants examined the association between eight voice appraisals and distress. Moderate to large summary effects (r ranging between 0.30 and 0.50) were observed in several analyses focusing on ‘maladaptive’ appraisals and beliefs about voices (malevolence, power, metaphysical beliefs, beliefs about loss of control, voice intrusiveness), with voice dominance having a large summary effect, r = 0.58, 95% CI [0.43, 0.69]. Positive appraisals and beliefs had small negative summary effects on distress. The magnitude of the observed effects was similar across subgroup analyses considering measures of voice-related distress, anxiety and depression. The findings of this evidence synthesis broadly corroborate cognitive-behavioural models of distressing voices, but suggested that factors other than voice appraisals may also predict the distress and impairment caused by hallucinatory experiences in people with schizophrenia-spectrum disorders. Nonetheless, our findings confirm that voice appraisals are an important and meaningful target for treatment in help-seeking voice hearers with psychosis.

Somatic and germline gain-of-function point mutations in RAF, the first oncogene to be discovered in humans, delineate a group of tumor-prone syndromes known as RASopathies. In this study, we document the first human phenotype resulting from the germline loss of function of the proto-oncogene RAF1 (a.k.a. CRAF). In a consanguineous family, we uncovered a homozygous p.Thr543Met mutation segregating with a neonatal lethal progeroid syndrome with cutaneous, craniofacial, cardiac and limb anomalies. Structure-based prediction and functional tests using human knock-in cells showed that threonine 543 is essential to: 1) ensure RAF1’s stability and phosphorylation, 2) maintain its kinase activity towards substrates of the MAPK pathway and 3) protect from stress-induced apoptosis. When injected in Xenopus embryos mutant RAF1T543M failed to phenocopy the effects of overactive FGF/MAPK signaling confirming its hypomorphic activity. Collectively, our data disclose the genetic and molecular etiology of a novel segmental progeroid syndrome which highlights the importance of RTK signaling for human development and homeostasis.Short summaryA germline homozygous recessive loss-of-function mutation p.T453M in RAF1 causes a neonatal lethal progeroid syndrome. In vitro and in vivo tests demonstrate that Thr543 is necessary for RAF1’s protein stability, to transduce signaling to the MAPK pathway and to respond to stress-induced apoptosis.

BACKGROUND Multiplex tests allow for measurement of allergen-specific IgE responses to multiple extracts and molecular allergens and have several advantages for large cohort studies. Due to significant methodological differences, test systems are difficult to integrate in meta-analyses/systematic reviews since there is a lack of datasets with direct comparison. We aimed to create models for statistical integration of allergen-specific IgE to peanut/tree nut allergens from three IgE test platforms. METHODS Plasma from Canadian and Austrian children/adolescents with peanut/tree nut sensitization and a cohort of sensitized, high-risk, pre-school asthmatics (total n = 166) were measured with three R&D multiplex IgE test platforms: Allergy Explorer version 1 (ALEX) (Macro Array Dx), MeDALL-chip (Mechanisms of Development of Allergy) (Thermo Fisher), and EUROLINE (EUROIMMUN). Skin prick test (n = 51) and ImmunoCAP (Thermo Fisher) (n = 62) results for extracts were available in a subset. Regression models (Multivariate Adaptive Regression Splines, local polynomial regression) were applied if >30% of samples were positive to the allergen. Intra-test correlations between PR-10 and nsLTP allergens were assessed. RESULTS Using two regression methods, we demonstrated the ability to model allergen-specific relationships with acceptable measures of fit (r2 = 94%-56%) for peanut and tree nut sIgE testing at the extract and molecular-level, in order from highest to lowest: Ara h 2, Ara h 6, Jug r 1, Ana o 3, Ara h 1, Jug r 2, and Cor a 9. CONCLUSION Our models support the notion that quantitative conversion is possible between sIgE multiplex platforms for extracts and molecular allergens and may provide options to aggregate data for future meta-analysis.

The 2,4-dinitrophenylhydrazine (2,4-DNPH) alpha-ketoglutarate detection assay was developed to support the study of prolyl hydroxylase domain (PHD) proteins in a substrate-independent manner. This protocol was extensively optimized for the PHD protein reaction, and is applicable to the study of enzyme kinetics or to high-throughput screening.

Cancer is a leading cause of morbidity and mortality, with up to 9.6 million cancer-related deaths worldwide in 2018 [1].

Numerous cancers, including prostate cancer (PCa), have been shown to be addicted to transcription programs driven by specific genomic sites known as superenhancers (SEs). Recently, it has been proposed that the robust transcription of genes at such SEs is enabled by the formation of phase-separated condensates by transcription factors and coactivators with intrinsically disordered regions. The androgen receptor (AR), the main oncogenic driver in PCa, contains large disordered regions and is co-recruited with the transcriptional coactivator MED1 to SEs in androgen-dependent prostate cancer cells, thereby promoting oncogenic transcriptional programs. In this work, we show that AR-rich, liquid-like foci form in prostate cancer models upon androgen stimulation. We reveal that foci formation correlates with AR transcriptional activity, which can be modulated by changing cellular foci content genetically or chemically. We also demonstrate that the transcriptional coactivator MED1 plays an essential role in the formation of transcriptionally active AR-rich foci and that AR antagonists that block cofactor recruitment or DNA binding hinder foci formation and thus AR transcriptional activity. The liquid like properties of the condensates were also validated in-vitro by using recombinant AR protein. Using clinical specimens, we detected the interaction between AR and MED1 in advanced forms of prostate cancer and not in benign tissues. These results suggest that enhanced compartmentalization of AR and coactivators at SEs may play an important role in the activation of oncogenic transcription programs in androgen-dependent PCa. A better understanding of the assembly and the regulation of these AR-rich compartments may provide novel therapeutic options for PCa by targeting downstream events of androgen activation and DNA binding of AR. Citation Format: Fan Zhang, Maitree Biswas, Joseph Lee, Shreyas Lingadahalli, Samantha Wong, Christopher Wells, Neetu Saxena, Bei Sun, Ana K. Parra-Nuñez, Christophe Sanchez, Jane Foo, Novia Chan, Lauren Ung, Nabeel Khan, Umut Berkay Altıntaş, Jennifer M. Bui, Yuzhuo Wang, Ladan Fazli, Paul S. Rennie, Nathan Lack, Artem Cherkasov, Martin E. Gleave, Joerg Gsponer, Nada Lallous. Dynamic phase separation of the androgen receptor and its coactivators to regulate gene expression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5729.

Climate change has been associated in phenological shifts for a variety of taxa. Amphibians, specifically the order Anura (frogs and toads), are considered particularly vulnerable due to their sensitivity to anthropogenic and environmental change. Previous research has documented shifts in the timing of anuran breeding that can be attributed, in part, to climate change, with potential implications for reproduction, survival, and development. This study aims to investigate how air temperature is associated with anuran calling phenology. I will examine the temporal trends in spring and summer air temperature in a lake in northern Ontario, Canada. and quantify seasonal patterns of calling anuran species using acoustic monitoring over a four-month period. I predict that there will be interspecific variation in peak calling associated with air temperature. Additionally, I expect to find asymmetrical association between air temperature and anuran species’ calling behaviour – wherein prolonged breeding species will have a larger optimal temperature range for calling compared to explosive breeding species. The findings of this research will aid in future conservation and provide insight for management strategies of anurans in Canada in response to anticipated climate warming.

Background Peanut and tree nut allergies are the most important causes of anaphylaxis. Co-reactivity to more than one nut is frequent, and co-sensitization in the absence of clinical data is often obtained. Confirmatory oral food challenges (OFCs) are inconsistently performed. Objective To investigate the utility of the basophil activation test (BAT) in diagnosing peanut and tree nut allergies. Methods The Markers Of Nut Allergy Study (MONAS) prospectively enrolled patients aged 0.5-17 years with confirmed peanut and/or tree nut (almond, cashew, hazelnut, pistachio, walnut) allergy or sensitization from Canadian (n = 150) and Austrian (n = 50) tertiary pediatric centers. BAT using %CD63+ basophils (SSClow/CCR3pos) as outcome was performed with whole blood samples stimulated with allergen extracts of each nut (0.001-1000 ng/mL protein). BAT results were assessed against confirmed allergic status in a blinded fashion to develop a generalizable statistical model for comparison to extract and marker allergen-specific IgE. Results A mixed effect model integrating BAT results for 10 and 100 ng/mL of peanut and individual tree nut extracts was optimal. The area under the ROC curve (AUROC) was 0.98 for peanut, 0.97 for cashew, 0.92 for hazelnut, 0.95 for pistachio, and 0.97 for walnut. The BAT outperformed sIgE testing for peanut or hazelnut and was comparable for walnut (AUROC 0.95, 0.94, 0.92) in a sub-analysis in sensitized patients undergoing OFC. Conclusions Basophil activation test can predict allergic clinical status to peanut and tree nuts in multi-nut-sensitized children and may reduce the need for high-risk OFCs in patients.

Objective This study explored the feasibility and acceptability of using Motivational Interviewing (MI) in the home setting with families of preschoolers. Methods Using mixed-methods pilot data from an MI-based obesity prevention intervention delivered via home visits by health educators (HEs) with 44 families (n = 17 four home visit group; n = 14 two home visit group), we examined: 1) fidelity of MI adherence by HEs; 2) parents’ perceptions of the intervention; and 3) HEs insights pertaining to the intervention’s delivery. Results Multiple measures of MI fidelity were deemed to exceed defined proficiency levels. Ninety-three percent of families reported being “satisfied” to “very satisfied” with the intervention. HEs reported building a high level of trust with families and gaining a thorough understanding of familial context. Parents appreciated how HEs’ were knowledgeable and provided personalized attention when discussing health goals. Some parents suggested more directive advice and follow-up visits as ways to improve the intervention. Conclusion Home-based MI was conducted with a high level of fidelity, was well accepted by families and practitioners. Practice implications Our findings from parents and MI practitioners provide key learnings that can inform future behavior change interventions that propose to use MI within the home setting.

It has been estimated that 1 in every 12 pupils is a young carer. Results from a Yorkshire secondary school survey, published here, show that this number could be even higher. This article describes how to work collaboratively to identify and support these vulnerable young people.

There is growing evidence of an association between the relative reinforcing value of food (RRVF; how hard a child is willing to work for a food versus a non-food reinforcer) and weight across many life stages, but few studies have examined this association among preschool age children. This study investigated the association between the RRVF (measured as the food reinforcing ratio (FRR)) and body mass index (BMI) z-score and percent fat mass (%FM), in 3 year-old children. The sample was comprised of 33 children who were 3 years of age (36–47 months). Children selected their favourite food reinforcer (cookie) and non-food (audiobook) reinforcer to be used in the FRR task, which was delivered using a sequential progressive fixed ratio reward schedule. Child height, weight and percent body fat using bioelectrical impedance analysis were assessed by trained research staff. Linear regression analyses showed that FRR was significantly and positively associated with BMI z-scores. FRR was not significantly associated with %FM. Findings from this study support the RRVF as a potential risk factor in the development of childhood obesity. Longitudinal research involving larger sample sizes would strengthen future research exploring associations between RRVF and body composition in preschool age children.

Carers are vital to the health economy, saving the NHS over £132 billion per year.1 This makes it fundamental to support them in both their carer and patient roles. A significant proportion of these carers are aged

Background: Childhood adversity, dissociation and adult attachment have all been implicated in the development of hallucinations or ‘voice-hearing’. Testing psychological models in relation to subclinical phenomena, such as proneness to hallucinations in non-clinical samples, provides a convenient methodology to develop understanding of the processes and mechanisms underlying clinical symptoms. Aims: This paper investigates the relative contribution of childhood adversity, dissociation and adult attachment in explaining hallucination proneness in a non-clinical sample. Methods: Students and staff with no previous contact with secondary care at the University of Manchester were recruited. Participants completed a series of self-report measures: the Launay‒Slade Hallucination Scale (LSHS), the Relationship Scale Questionnaire (RSQ), the Childhood Trauma Questionnaire (CTQ), the Dissociative Experiences Schedule (DES II) and the Positive and Negative Affect Schedule (PANAS). Results: As hypothesized, insecure attachment, childhood adversity and dissociative symptoms were correlated with hallucination proneness. Multiple regression analysis, controlling for confounds of age and negative affect, indicated that the RSQ, CTQ and DES II predicted hallucination proneness. Only DES II and RSQ avoidant attachment were significant independent predictors in the final model. Conclusions: This study provides further evidence to support the idea that attachment and dissociation are important psychological mechanisms involved in voice-hearing proneness. Further testing is required with a clinical population.

Non-clinical participants viewed pictures of either neutral or stressful interpersonal scenes before completing a voice-detection task. Exposure to stress did not influence voice detection, but scores on a measure of depersonalisation were positively correlated with voices.

The aim was to assess in vitro the antibacterial activity of 10 root canal lubricants. K-Y Jelly personal lubricant, RC-Prep, File-Eze, File-Rite, EndoPrep Gel, Endosure Prep Creme 15%, Prep-Rite, Glyde, SlickGel ES and Alpha Glide were selected and compared in their antimicrobial properties to seven irrigants. Serial dilutions of each agent in tryptic soy broth were inoculated with either Enterococcus faecalis or Pseudomonas aeruginosa and incubated at 37C for 24 h. During incubation bacterial growth was measured by optical density (A(600)), and samples removed for cultivation on tryptic soy broth agar. Against both test bacteria after 1 h incubation, six lubricants recorded minimum bactericidal concentrations ranging from 1/10 to 1/80, whereas the inhibitory activity of the irrigants ranged from 1/20 to 1/640. Under these conditions, several lubricants exhibited antimicrobial activity comparable with some irrigants. Three irrigants, Consepsis (containing chlorhexidine), Endosure EDTA/C (containing cetrimide) and EndoPrep Solution (containing cetrimide), showed superior antibacterial action to lubricants against both species. The irrigants containing ethylenediamine tetraacetic acid and cetrimide were the most effective against both bacterial species at all time intervals. Antimicrobial activity of the lubricants did not correlate to pH values, which ranged from 2.9 to 10.3. Root canal lubricants have antibacterial properties that may help to disinfect canals.

BACKGROUND: Understanding the information needs and preferred vehicles of information delivery to patients with inflammatory bowel disease (IBD) will enhance their care.OBJECTIVE: To survey persons with longstanding IBD as to their information needs and preferred vehicles of information delivery.METHODS: The population-based Manitoba IBD Cohort (n=271, mean disease duration 11 years) was surveyed to assess its information needs across 23 issues, both retrospectively at the time of diagnosis and currently.RESULTS: Most participants (64%) were initially diagnosed by a gastroenterologist, or otherwise by a family physician (19%) or surgeon (12%). Recalling time of diagnosis, at least 80% rated as very important information about common symptoms of IBD, possible complications, long-term prognosis, medication side effects, self management of symptoms and when to involve the doctor, yet only 10% to 36% believed they received the right amount of information about these issues. Dietary guidance was also regarded as important by 80% to 89%, yet only 8% to 16% received the correct amount of information. Regarding current needs, a large proportion believed it would be very helpful to have more information about long-term prognosis (66%) and diet considerations (60% to 68%). The following information sources were regarded as very acceptable: medical specialist (81%); brochure (79%); family doctor (64%); and website (64%), with 51% ranking the medical specialist as the first choice. In a comparison of the responses of this cohort to those of a recently diagnosed sample, there was remarkable consistency in the information needs and most desired sources of information.DISCUSSION: In the present population-based cohort with longstanding disease, dietary information was regarded as the least adequately addressed. There was clear openness to receiving information through other routes than just the medical specialist, suggesting that optimizing brochures and websites would be an important adjunct source of information.CONCLUSION: Approximately 10 years after diagnosis, only a small percentage of persons with IBD believed they received the correct amount of information about the issues they regarded as most important to have discussed at diagnosis.

Objectives: The objectives of the study were: (i) to survey staff perceptions of causes of delay in patients' journeys through the ED; (ii) to identify and analyse key constraints to patient flow using real-time diagnostic/decision support software (Patient Flow Study); and (iii) to assess the correlation between staff perceptions and data from the Patient Flow Study. Methods: ED and non-ED staff were surveyed prior to the Patient Flow Study. The survey involved ranking the likely reasons for delay at three set points after triage (160, 320 and 480 min). Real-time data on delay in patients' journeys through the ED were collected over a period of 5 weeks. The correlation between staff perceptions and study data at the three time points was calculated using Spearman's rank correlation coefficient. Subgroup analysis was performed on the basis of staff position, years of experience at St George and whether they had previously attended training on constraints to flow. Results: A total of 68 staff responded to the questionnaire (response rate 42%). During the study period, 4555 ED attendances (97% of all presentations) were analysed for causes of constraint. Strong correlation between staff perceptions and real-time data was only found among some subgroups at the point 160 min from triage. Conclusions: Overall, staff perceptions regarding causes of constraint to patient flow do not correlate well with data obtained from real-time analysis.

During latency, Epstein-Barr virus (EBV) is stably maintained as a circular plasmid that is replicated once per cell cycle and partitioned at mitosis. Both these processes require a single viral protein, EBV nuclear antigen 1 (EBNA1), which binds two clusters of cognate binding sites within the latent viral origin, oriP . EBNA1 is known to associate with cellular metaphase chromosomes through chromosome-binding domains within its amino terminus, an association that we have determined to be required not only for the partitioning of oriP plasmids but also for their replication. One of the chromosome-binding domains of EBNA1 associates with a cellular nucleolar protein, EBP2, and it has been proposed that this interaction underlies that ability of EBNA1 to bind metaphase chromosomes. Here we demonstrate that EBNA1's chromosome-binding domains are AT hooks, a DNA-binding motif found in a family of proteins that bind the scaffold-associated regions on metaphase chromosomes. Further, we demonstrate that the ability of EBNA1 to stably replicate and partition oriP plasmids correlates with its AT hook activity and not its association with EBP2. Finally, we examine the contributions of EBP2 toward the ability of EBNA1 to associate with metaphase chromosomes in human cells, as well as support the replication and partitioning of oriP plasmids in human cells. Our results indicate that it is unlikely that EBP2 directly mediates these activities of EBNA1 in human cells.

The aim was to assess in vitro the antibacterial activity of 10 root canal lubricants. K-Y Jelly personal lubricant, RC-Prep, File-Eze, File-Rite, EndoPrep Gel, Endosure Prep Crème 15%, Prep-Rite, Glyde, SlickGel ES and Alpha Glide were selected and compared in their antimicrobial properties to seven irrigants. Serial dilutions of each agent in tryptic soy broth were inoculated with either Enterococcus faecalis or Pseudomonas aeruginosa and incubated at 37C for 24 h. During incubation bacterial growth was measured by optical density (A(600)), and samples removed for cultivation on tryptic soy broth agar. Against both test bacteria after 1 h incubation, six lubricants recorded minimum bactericidal concentrations ranging from 1/10 to 1/80, whereas the inhibitory activity of the irrigants ranged from 1/20 to 1/640. Under these conditions, several lubricants exhibited antimicrobial activity comparable with some irrigants. Three irrigants, Consepsis (containing chlorhexidine), Endosure EDTA/C (containing cetrimide) and EndoPrep Solution (containing cetrimide), showed superior antibacterial action to lubricants against both species. The irrigants containing ethylenediamine tetraacetic acid and cetrimide were the most effective against both bacterial species at all time intervals. Antimicrobial activity of the lubricants did not correlate to pH values, which ranged from 2.9 to 10.3. Root canal lubricants have antibacterial properties that may help to disinfect canals.

Essential hypertension often requires affected patients to self-manage their condition most of the time. Besides seeking regular medical review of their life-long condition to detect vascular complications, patients have to maintain healthy lifestyles in between physician consultations via diet and physical activity, and to take their medications according to their prescriptions. Their self-management ability is influenced by their self-efficacy capacity, which can be assessed using questionnaire-based tools. The "Hypertension Self-Care Profile" (HTN-SCP) is 1 such questionnaire assessing self-efficacy in the domains of "behavior," "motivation," and "self-efficacy." This study aims to determine the test-retest reliability of HTN-SCP in an English-literate Asian population using a web-based approach. Multiethnic Asian patients, aged 40 years and older, with essential hypertension were recruited from a typical public primary care clinic in Singapore. The investigators guided the patients to fill up the web-based 60-item HTN-SCP in English using a tablet or smartphone on the first visit and refilled the instrument 2 weeks later in the retest. Internal consistency and test-retest reliability were evaluated using Cronbach's Alpha and intraclass correlation coefficients (ICC), respectively. The t test was used to determine the relationship between the overall HTN-SCP scores of the patients and their self-reported self-management activities. A total of 160 patients completed the HTN-SCP during the initial test, from which 71 test-retest responses were completed. No floor or ceiling effect was found for the scores for the 3 subscales. Cronbach's Alpha coefficients were 0.857, 0.948, and 0.931 for "behavior," "motivation," and "self-efficacy" domains respectively, indicating high internal consistency. The item-total correlation ranges for the 3 scales were from 0.105 to 0.656 for Behavior, 0.401 to 0.808 for Motivation, 0.349 to 0.789 for Self-efficacy. The corresponding ICC scores of 0.671, 0.762, and 0.720 for these respective domains showed good test-retest reliability. The correlation of the HTN-SCP scores and patients' reported self-management measures were significant, except for keeping their food diary. HTN-SCP showed satisfactory internal consistency and test-retest reliability in an English literate Asian population. A web-based approach is feasible if similar studies are needed to validate its translated versions of the tool for wider application in the local multilingual population.

Peak force (PF), contractile rate of force development (RFD) and contractile impulse (CI) are of great importance to competitive weightlifters (WL). These athletes routinely perform successive bouts of high-intensity resistance exercise (HIRE) within the same day (double-day training) with the aim of improving muscular function and weightlifting performance. The purpose of this investigation was to determine and compare the PF, contractile RFD and CI responses to double-day training between WL and resistance trained (RT) adults (n = 16 per group). Furthermore, we sought to establish whether acute changes in muscle function were associated with acute changes in muscle architecture. Isometric front squat PF, contractile RFD, CI and the pennation angle (θ(p)), anatomical and physiological thickness of the m. vastus lateralis (VL) were determined before and after two equivalent HIRE sessions separated by 4-6 h rest. Each session consisted of ten single repetitions of the dynamic barbell front squat interspersed with 2-min rest, using a load equivalent to 90% of the pre-session PF. Weightlifters demonstrated greater PF at all time points when compared to RT adults and exhibited no significant within or between session changes in PF, contractile RFD or CI. Conversely, RT adults demonstrated within- and between-session decreases in PF and between-session increases in contractile RFD and CI. As no correlations were found between the relative within-session changes in muscle function and the concomitant changes in muscle architecture, other factors must contribute to the divergent responses in PF, contractile RFD and CI between WL and RT adults.